[OpenSPIM] About the calibration of the lightsheet

Wed Aug 7 07:00:14 CDT 2013

 Hi Jan,

Thanks a lot for your comments. 
I will search in the lab if there is a little mirror, otherwise I should buy it.
The method that you used is also very clever. That will be the one that I will
use since looks like it is cheaper to buy a little simple mirror.

Kind regards,
Edgar

 _________________________________________________________________________

Fraunhofer-Institut für Produktionstechnologie IPT 
Edgar Escobar Nieto  

Steinbachstraße 17 
52074 Aachen 

edgar.escobar.nieto at ipt.fraunhofer.de 
http://www.ipt.fraunhofer.de
_________________________________________________________________________

-----"Krieger, Jan" <j.krieger at Dkfz-Heidelberg.de> schrieb: -----
An: "edgar.escobar.nieto at ipt.fraunhofer.de" <edgar.escobar.nieto at ipt.fraunhofer.de>
Von: "Krieger, Jan" <j.krieger at Dkfz-Heidelberg.de>
Datum: 07.08.2013 12:21
Betreff: AW: [OpenSPIM] About the calibration of the lightsheet

Hi!

we are using a simple but small silver mirror (5x5mm², 1mm thick, bought from Melles-Griot) on a special mount, but you can glue it to anything that can be mounted inside the SPIM. First we focus the mirror, by illuminating it with a white LED or sinple lamp through the projection objective. The mirror is small enough to also fit into an openSPIM (I did that together with Alexis Maizel two weeks ago). Basically you can focus onto some dirk on the mirror surface, or a skratch that we make into it. Then you can switch to the laser and first bring it to the sharp line on the mirror and finally align it with the lenses of the illumination beam path. I think this is kind of the "classical" method to do that. 

The samples seem to be embedded in glass, right? Then you can focus them, but you won't have a focussed reflective surface then, that you can use to inspect the lightsheet.

Best,
JAN

Dipl.-Phys. Jan Krieger
German Cancer Research Center (dkfz)
Department B040 - Biophysics of Macromolecules (Prof. J. Langowski)
Im Neuenheimer Feld 580
69120 Heidelberg

fon: +49 / 6221 / 42-3395
fax: +49 / 6221 / 42-3391
e-mail: j.krieger at dkfz.de
www: http://www.dkfz.de/Macromol/
________________________________________
Von: openspim-bounces at openspim.org [openspim-bounces at openspim.org] im Auftrag von edgar.escobar.nieto at ipt.fraunhofer.de [edgar.escobar.nieto at ipt.fraunhofer.de]
Gesendet: Mittwoch, 7. August 2013 12:13
An: Peter Gabriel Pitrone
Cc: openspim at openspim.org
Betreff: Re: [OpenSPIM] About the calibration of the lightsheet

Hi dear Peter,

I have not yet bought the Ronchi Ruler Slide, but I was thinking of use a different
material in order to perform the alignment of the lighstsheet. I was thinking of use
some fluorescence samples that are mounted in a glass http://www.bw-optik.de/laufband/fluoreszenz.php
and since I have already some of this mounted glass samples available I will try to make a similar procedure as the stated in the OpenSPIM wiki.
If that method doesn't work I will have to buy the Ronchi Ruler Slide anyway.

The think is that since the material is also glass, would be the same to cut it or to cut the Ronchi Ruler Slide. That's
why I was asking how the Ruler Slide was cut. I will try to find a way to cut the glass, the problem
is that only a thin piece, let's say from 1 to 2 mm wide is needed. I know that would be so difficul to perform a cut
of this magnitude using a conventional glass cutter, but I'll give a try, if I am successful I will write a protocol to the OpenSPIM wiki.

I think until I had all the necessary material ready and mounted and if I have troubles with the alignment of the lightsheet,
that would be nice to have the Skype session.

Thanks for your kind answer and for offering me your support. Enjoy your holidays in the U.S.

Kind regards,
Edgar
_________________________________________________________________________

Fraunhofer-Institut für Produktionstechnologie IPT
Edgar Escobar Nieto

Steinbachstraße 17
52074 Aachen

edgar.escobar.nieto at ipt.fraunhofer.de
http://www.ipt.fraunhofer.de <http://www.ipt.fraunhofer.de/>
_________________________________________________________________________

-----"Peter Gabriel Pitrone" <pitrone at mpi-cbg.de> schrieb: -----
An: edgar.escobar.nieto at ipt.fraunhofer.de
Von: "Peter Gabriel Pitrone" <pitrone at mpi-cbg.de>
Datum: 06.08.2013 18:22
Kopie: openspim at openspim.org
Betreff: Re: [OpenSPIM] About the calibration of the lightsheet

Hello Edgar,

I'm sorry I didn't answer you till now, but I'm sometimes away from my
email as I am on vacation in the states.

To answer your questions, we broke the slide and found the best shard. It
is definitely not the most elegant way, but it works for us. If you want
to go about making it with more consistency with a glass or window
cutter, please write a protocol on our wiki on how to do it so others can
reproduce it.

If you need help with aligning the light sheet, we can make a skype
session tomorrow... just remember I have an 8 hour difference in
timezones. My Skype name is "petepitrone". send me an email as to the
time you would like to talk.

Pete

--
Peter Gabriel Pitrone - TechRMS
Microscopy/Imaging Specialist
Prof. Dr. Pavel Tomancak group
Max Planck Institute for
Molecular Biology and Genetics
Pfotenhauerstr. 108
01307 Dresden

"If a straight line fit is required, obtain only two data points." - Anon.

On Tue, August 6, 2013 5:08 pm, edgar.escobar.nieto at ipt.fraunhofer.de wrote:
<|>  Hi dear All,
<|>
<|> I would like to know how you could cut the ruling slides. I don't
know if
<|> it is possible tu cut a thin part of glass using
<|> a conventional glass cutter.
<|>
<|> Kind regards,
<|> Edgar Escobar Nieto
<|>
_________________________________________________________________________
<|>
<|> Fraunhofer-Institut fr Produktionstechnologie IPT
<|> Edgar Escobar Nieto
<|>
<|>
<|>
<|>
<|> Steinbachstraáe 17
<|> 52074 Aachen
<|>
<|> edgar.escobar.nieto at ipt.fraunhofer.de
<|> http://www.ipt.fraunhofer.de
<|>
_________________________________________________________________________
<|>
<|>
<|>
<|>
<|>
<|> -----Weitergeleitet von Edgar Escobar Nieto/Fraunhofer IPT am 06.08.2013
<|> 17:01 -----
<|> An: openspim at openspim.org
<|> Von: Edgar Escobar Nieto/Fraunhofer IPT
<|> Datum: 05.08.2013 11:55
<|> Betreff: About the calibration of the lightsheet
<|>
<|>
<|>
_________________________________________________________________________
<|>
<|> Fraunhofer-Institut fr Produktionstechnologie IPT
<|> Edgar Escobar Nieto
<|>
<|>
<|>
<|>
<|> Steinbachstraáe 17
<|> 52074 Aachen
<|>
<|> edgar.escobar.nieto at ipt.fraunhofer.de
<|> http://www.ipt.fraunhofer.de
<|>
_________________________________________________________________________
<|>
<|>
<|>
<|>
<|>
<|> -----Weitergeleitet von Edgar Escobar Nieto/Fraunhofer IPT am 05.08.2013
<|> 11:54 -----
<|> An: openspim at openspim.org
<|> Von: Edgar Escobar Nieto/Fraunhofer IPT
<|> Datum: 31.07.2013 15:02
<|> Betreff: (Unbenannt)
<|>
<|> ÿHi dear all,
<|>
<|> I reached to the point where I need to calibrate the light-sheet. And I
<|> was wondering
<|> what is the optical density of the ND filter recommended to perform the
<|> calibration.
<|>
<|> I would like to know also what is the Frequency (lp/mm) recommended for
<|> the Opal Glass Ronchi Ruling Slides.
<|> I think that high a frequency would be better, but there must be a
limit.
<|>
<|> Only to be sure, what I understood is that these Ruling Slides have
<|> already a reflective surface, so there is no
<|> need to glue a mirror to the ruling slides.
<|>
<|> Thank you in advance for your comments.
<|>
<|> Kind regards,
<|> Edgar Escobar Nieto
<|>
_________________________________________________________________________
<|>
<|> Fraunhofer-Institut fr Produktionstechnologie IPT
<|> Edgar Escobar Nieto
<|>
<|>
<|>
<|>
<|> Steinbachstraáe 17
<|> 52074 Aachen
<|>
<|> edgar.escobar.nieto at ipt.fraunhofer.de
<|> http://www.ipt.fraunhofer.de
<|>
_________________________________________________________________________
<|>
<|>
<|>
<|>
<|>
<|> _______________________________________________
<|> OpenSPIM mailing list
<|> OpenSPIM at openspim.org
<|> http://openspim.org/mailman/listinfo/openspim
<|>

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